Related gene for regulating and controlling wood development and application thereof

A gene and exogenous gene technology, applied in the field of biotechnology and bioenergy, can solve the problems of large environmental pollution

Active Publication Date: 2016-08-03
CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional bleaching agents are mostly chlorine-containing bleaching agents. Wastewater contains toxic substances with strong carcinogenicity, which pollutes the environment greatly.
[0007] Therefore, there is no wood plant suitable for the production of biofuels and papermaking in this field. Therefore, it is urgent to study the genes related to the regulation of wood development and develop wood species suitable for the production of renewable energy and papermaking.

Method used

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  • Related gene for regulating and controlling wood development and application thereof
  • Related gene for regulating and controlling wood development and application thereof
  • Related gene for regulating and controlling wood development and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0168] The cloning of the gene expression sequence of embodiment 1PtMAN6 and promoter

[0169] 1. Sequence information

[0170] The complete CDS sequence of PtMAN6 is as follows:

[0171]

[0172] or

[0173]

[0174]

[0175] The amino acid sequence of PtMAN6 protein is as follows:

[0176]

[0177] or

[0178]

[0179] The cloned PtMAN6 promoter sequence is as follows:

[0180]

[0181]

[0182] 2. Vector construction

[0183] 2.1 Overexpression vector

[0184] 35S:PtMAN6 vector map see figure 1 ; 35S:PtMAN6-GFP vector map figure 2 .

[0185] 2.2 Antisense expression vector

[0186] 35S:PtMAN6AS map see image 3 .

[0187] 2.3 Transient expression vector

[0188] MAN6G transient expression vector see Figure 4 , MAN6sG transient expression vector see Figure 5 .

Embodiment 2

[0189]Example 2 Overexpression of PtMAN6 reduces the content of wood lignin and crystalline cellulose

[0190] Western detection proves that the PtMAN6 overexpression (OE) vector and antisense (AS) expression vector are successfully transferred into poplar, and can be expressed in plants, and further play a role ( Figure 6 A).

[0191] Analysis of transgenic plants showed that compared with wild-type plants, the petioles and stems of OE plants became softer ( Figure 6 B, 6C and 6D), while AS plants were the opposite ( Figure 6 B, 6C and 6E), indicating that PtMAN6 is involved in the regulation of stem mechanical strength. Further slice analysis showed that the content of lignin in the petiole of OE plants decreased ( Figure 6 F-H). Similarly, lignin deposition in stems also decreased ( Figure 7 A-I). The determination of lignin and crystalline cellulose content of mature wood further proved the function of PtMAN6, and overexpression of this gene resulted in a decrea...

Embodiment 3

[0192] The biomass of embodiment 3 transgenic poplar does not change significantly

[0193] The present embodiment has measured the biomass of poplar, and the result shows, with respect to wild type, the height and girth of 1.5 year old transgenic poplar wood all have no significant change ( Figure 8 ).

[0194] Figure 8 Shows the height and girth analysis results of 1.5-year-old transgenic poplar wood, wherein, Figure 8 A shows the overexpression of PtMAN6 poplar (OE), the antisense expression of PtMAN6 poplar (AS) and the wild type (WT) poplar have no significant difference in trunk height; Figure 8 B shows that there is no significant difference in the circumference of the base of the trunk of OE, AS and WT poplar, and the statistical analysis uses T test.

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Abstract

The invention relates to a related gene for regulating and controlling wood development and application thereof, and particularly provides a novel PtMAN6 polypeptide or a coding sequence thereof. An amino acid sequence of the PtMAN6 polypeptide is shown as SEQ ID NO.:2, and the coding sequence is shown as SEQ ID NO.:1.The invention further provides a PtMAN6 gene and application of protein coded by the PtMAN6 gene. The PtMAN6 gene can reduce synthesis of xylem secondary cell walls and / or reduce accumulation of crystalline fiber biomass; mannan hydrolase of mannan hemicellulose can be enriched and degraded in plants by excessively expressing the PtMAN6; meanwhile, due to the regulating and controlling effect of the PtMAN6, the lignin content in transgenic plants is decreased, crystalline cellulose is decreased, and therefore the cost in lignocelluloses ethanol producing and paper making industry is greatly reduced; a promoter of the PtMAN6 gene has the specific duct cell positioning function and can precisely regulate and control expression of the gene in xylem duct cells.

Description

[0001] This application is a divisional application of an invention application with an application date of June 1, 2012, an application number of 201210179885.7, and an invention title of "A Gene Regulating Wood Development and Its Application". technical field [0002] The invention belongs to the fields of biotechnology and bioenergy, in particular, the invention relates to a gene regulating wood development and its application. Background technique [0003] With the development of social economy and the continuous increase of population, human beings demand more and more energy. Due to the limited fossil energy, the prices of petroleum and other products continue to rise, which poses a threat to the stable and sustainable development of the economy. [0004] Renewable biomass energy is one of the ideal alternative energy sources, and bioethanol is currently the most promising and main biomass energy source. Since the oil crisis in the mid-1970s, some countries, mainly th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/82C12N9/42C12N15/56A01H5/00
CPCC12N9/2494C12N15/8226C12Y302/01078
Inventor 李来庚赵运军
Owner CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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